• Title/Summary/Keyword: DNA-based testing

Search Result 95, Processing Time 0.026 seconds

Molecular Systematics of Tephritidae (Insecta : Diptera): Testing Phylogenetic Position of Korean Acidiella spp. (Trypetini) Using Mitochondrial 16S rDNA Sequences

  • Han, Ho-Yeon;Ro, Kyung-Eui
    • Animal cells and systems
    • /
    • v.6 no.1
    • /
    • pp.13-18
    • /
    • 2002
  • Phylogenetic relationships of Korean Acidiella species were tested using mitochondrial 16S ribosomal RNA gene sequences. We used 16 published sequences as outgroup, and 10 new sequences for nine Korean Acidiella species as ingroup. The number of aligned sites was 1,281 bp, but 1,135 bp were used for the analysis after excluding sites with missing data or gaps. Among these 1,135 sites, 464 sites were variable and 340 were informative for parsimony analysis. Phylogenetic information was extracted from this data set using neighbor-joining, maximum likelihood and maximum parsimony methods and compared to a morphology-based phylogenetic hypothesis. Our molecular data suggest that: (1) the tribe Trypetini appears to be monophyletic even when the nine additional Acidiella species are added to our previous phylogenetic analysis; (2) all the Korean Acidiella species belong to the Trypeta group, but the genus Acidiella is not supported as monophyletic; (3) the close relationship of A. circumvaga, A. issikii, and A. sapporensis is supported; (4) the close relationship of A. pachypogon and two additional new Acidiella species is strongly supported; and (5) the possible presence of two or more cryptic species among the specimens previously identified as A. obscuripennis is suggested. Sequence data from the mitochondrial 16S rDNA allowed us to better understand the systematic status of Korean Acidiella species. They indicated that the current concept about the genus Acidiella is insufficient and needs to be refined further. This study also showed a few interesting relationships, that had not been recognized by morphological study alone. Based on this study, we were able to plan further experiments to analyze relationships within the Trypeta Group.

Hybrid Capture 2 Assay Based Evaluation of High-Risk HPV Status in Healthy Women of North-East India

  • Rai, Avdhesh Kumar;Das, Debabrata;Kataki, Amal Chandra;Barmon, Debabrata;Aggarwal, Dippy;Deka, Pankaj;Shrivastava, Sushruta;Sharma, Jagannath Dev;Sarma, Anupam;Baruah, Upasana;Sharma, Mousumi
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.2
    • /
    • pp.861-865
    • /
    • 2014
  • Background: High risk HPV (HR-HPV) testing has been recommended as an effective tool along with cytology screening in identification of cervical intraepithelial lesions (CINs) and prevention of their progress towards invasive cervical cancer. The aim of this study was to assess the HR-HPV DNA status by Hybrid Capture 2 (HC2) assay in healthy asymptomatic women of North-East India. Materials and Methods: This study examined cervical cell samples of forty three (n=43) healthy women by HC2 assay. A High Risk HPV DNA kit (Qiagen) was used which can detect 13 high risk HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68. Results: The mean relative light units (RLU) for samples was in the range of 141-5, 94, 619. HR-HPV DNA was confirmed in 16% (7/43) of participant women samples. Among demographic and clinical parameters, menstrual irregularity (p=0.039) and infection history (p=0.028) has shown statistically significant differences between the HR-HPV-positive and negative groups. In the HR-HPV positive group, two women were confirmed for CINs after colposcopy and histopathologic examination. Conclusions: We suggest that there may be an association between irregular menstruation and infection history of the urogenital tract with HR-HPV DNA prevalence in North-East Indian asymptomatic women. HC2 assay can be a valuable tool for HR-HPV screening.

Effects of Rhodobacter sp. SA16 on Lettuce(Lactuca sativa L.) in Plastic Film House (시설 상추에 대한 Rhodobacter sp. SA16 처리 효과)

  • Lee, Young-Han;Jeong, Han-Taek;Yun, Han-Dae
    • Korean Journal of Environmental Agriculture
    • /
    • v.27 no.2
    • /
    • pp.163-170
    • /
    • 2008
  • This experiment was conducted to determine the development of mixed organic fertilizer using photosynthetic bacteria and mass production of mixed microbial compound for the environment-friendly agriculture. Photosynthetic bacteria, Rhodobacter sp. SA16 was isolated from soil collected by plastic film house. The SA16 strain was identified based on 16S rDNA sequence analysis and it is closely related to Rhodobacter sp.(100% similarity). The mixed organic fertilizer using SA16 was made of $N-P_2O_5-K_2O=60-10-20\;g\;kg^{-1}$ with combined soybean cake, sesame cake, powdered blood, fish meal, powdered bones and red-yellow soil. The mixed organic fertilizer 0.45, 0.90 and 1.35 Mg $ha^{-1}$ application in Ihyeon series was treated based on soil testing for lettuce cultivation in plastic film house. These results showed that the yield was increased the 18 and 19%over control by the mixed organic fertilizer application 0.45 and 0.90 Mg $ha^{-1}$, respectively. In the physical properties of the soil, the porosity of mixed organic fertilizer 1.35 Mg $ha^{-1}$ treatment was highest at 58.8%. Our results clearly revealed that the organic fertilizer using Rhodobacter sp. SA16 and mass production of mixed strains could be a useful technology in pursuing environment-friendly agriculture.

Evaluation of the Genetic Toxicity of Synthetic Chemicals (XIII) - Single Cell Gel Electrophoresis of Benzoyl Chloride, 2-Propyn-1-ol, and 2-Phenoxyethanol in Chinese Hamster lung Fibroblast -

  • Ryu, Jae-Chun;Kim, Youn-Jung
    • Environmental Mutagens and Carcinogens
    • /
    • v.24 no.2
    • /
    • pp.79-84
    • /
    • 2004
  • Three synthetic chemicals, benzoyl chloride, 2-propyn-l-ol, and 2-phenoxy ethanol were selected for genotoxicity testing, based on production quantity and available genotoxic data. In our previous report, benzoyl chloride induced chromosomal aberrations in Chinese hamster lung (CHL) fibroblast in vitro with and without metabolic activation, while 2-propyn-l-ol and 2-phenoxy ethanol induced only with metabolic activation. To compare the genotoxicity of chromosome aberration assay, the single cell gel electrophoresis (comet) assay subjected using CHL cells. As a result, statistically significant differences of tail moment values of benzoyl chloride, 2-propyn-1-ol, and 2-phenoxy ethanol were observed compared with control values on almost all concentrations with S9 or without S9 metabolic activation system. This results suggest that genotoxic results of the comet assay and the chromosome aberration assay show correlationship of genotoxicity in the CHL fibroblast. In summary, the positive result of chromosome aberration of benzoyl chloride, 2-propyn-l-ol, and 2-phenoxy ethanol was also induced DNA damages in comet assay with same cell line. Consequently, comet assay will be useful and more accurate tool to detect and to confirm the genotoxicity especially DNA damages in CHL fibroblast.

  • PDF

Application of digital polymerase chain reaction technology for noninvasive prenatal test

  • Lee, Seung Yong;Hwang, Seung Yong
    • Journal of Genetic Medicine
    • /
    • v.12 no.2
    • /
    • pp.72-78
    • /
    • 2015
  • Recently, noninvasive prenatal test (NIPT) has been adopted as a primary screening tool for fetal chromosomal aneuploidy. The principle of NIPT lies in isolating the fetal fraction of cell-free DNA in maternal plasma and analyzing it with bioinformatic tools to measure the amount of gene from the target chromosome, such as chromosomes 21, 18, and 13. NIPT will contribute to decreasing the need for unnecessary invasive procedures, including amniocentesis and chorionic villi sampling, for confirming fetal aneuploidy because of its higher positive predictive value than that of the conventional prenatal screening method. However, its greater cost than that of the current antenatal screening protocol may be an obstacle to the adoption of this innovative technique in clinical practice. Digital polymerase chain reaction (dPCR) is a novel approach for detecting and quantifying nucleic acid. dPCR provides real-time diagnostic advantages with higher sensitivity, accuracy, and absolute quantification than conventional quantitative PCR. Since the groundbreaking discovery that fetal cell-free nucleic acid exists in maternal plasma was reported, dPCR has been used for the quantification of fetal DNA and for screening for fetal aneuploidy. It has been suggested that dPCR will decrease the cost by targeting specific sequences in the target chromosome, and dPCR-based noninvasive testing will facilitate progress toward the implementation of a noninvasive approach for screening for trisomy 21, 18, and 13. In this review, we highlight the principle of dPCR and discuss its future implications in clinical practice.

Distribution of Human Papilloma Virus Infections of Uterine Cervix among Women of Reproductive Age - a Cross Sectional Hospital-Based Study from North East India

  • Sarma, Usha;Mahanta, Jagadish;Borkakoty, Biswajyoti;Sarmah, Bidula
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.16 no.4
    • /
    • pp.1519-1523
    • /
    • 2015
  • Infection of the uterine cervix by human papilloma viruses (HPV) may be associated with cervical pre-cancer and invasive cervical carcinoma if left untreated. With advance in molecular techniques, it has become easier to detect the resence of HPV DNA long before the appearance of any lesion. This study concerned cervical scrape samples of 310 married non-pregnant women attending a gynecology outpatient department for both Pap and PCR testing to detect HPV DNA. Nested PCR using primers for L1 consensus gene with My9/My11 and GP6+/GP5+followed by multiplex PCR were carried out to detect HPV 16 and HPV18. Result: HPV prevalence was 11.9% out of which 3.67% cases of negative for intra-epithelial lesion or malignancy (NILM) and in 71.1% (27/38) of atypical cervical smears were HPV positive. There was increasing trend of high-risk-HPV positivity (HR HPV 16 and 18), from 20% in benign cytology (NILM) to 42.9 % in LSIL, 71.41% in HSIL and 100% in SCC. There was highly significant association of HPV infection with cervical lesion ($x^2=144.0$, p<0.01) and also with type specific HPV prevalence ($x^2=7.761^*$, p<0.05).

Development of Sequence-Based DNA Markers for Evaluation of Phylogenetic Relationships in Korean Watermelon Varieties

  • Lee, Hee-Jeong;Cho, Hwa-Jin;Lee, Kyung-Ah;Lee, Min-Seon;Shin, Yoon-Seob;Harn, Chee-Hark;Yang, Seung-Gyun;Nahm, Seok-Hyeon
    • Journal of Crop Science and Biotechnology
    • /
    • v.10 no.2
    • /
    • pp.98-105
    • /
    • 2007
  • Phylogenetic relationships in Korean watermelons were evaluated by genetic similarity coefficients using 15 SSR(simple sequence repeat), 14 SCAR(sequence characterized amplified region) and 14 CAPS(sequence characterized amplified region) markers. The SSR markers were selected from previously reported melon and watermelon SSRs through testing polymorphisms within a set of commercial $F_1$ varieties. The SCAR and CAPS markers were developed from polymorphic AFLP(amplified fragment length polymorphism) markers between inbred lines 'BN4001' and 'BN4002'. From the AFLP analysis, 105 polymorphic fragments were identified between the inbred lines using 1,440 primer combinations of EcoRI+CNNN and XbaI+ANNN. Based on the sequencing data of these polymorphic fragments, we synthesized sequence specific primer pairs and detected clear and reliable polymorphisms in 27 primer pairs by indels(insertion/deletion) or RFLP(restriction fragment length polymorphism). A total of 43 sequence-based PCR markers were obtained and polymorphic information content(PIC) was analyzed to measure the informativeness of each marker in watermelon varieties. The average PIC value of SCAR markers was 0.41, which was similar to that of SSR markers. Genetic diversity was also estimated by using these markers to assess the phylogenetic relationships among commercial varieties of watermelon. These markers differentiated 26 Korean watermelon varieties into two major phylogenetic groups, but this grouping was not significantly correlated with their morphological and physiological characteristics. The mean genetic similarity was 66% within the complete set of 26 commercial varieties. In addition, these sequence-based PCR markers were reliable and useful to identify cultivars and genotypes of watermelon.

  • PDF

Prevalence of Dirofilaria immitis Infection in Stray Cats by Nested PCR in Korea

  • Park, Hyung-Jin;Lee, Sang-Eun;Lee, Won-Ja;Oh, Jung-Hyun;Maheswaran, Easwaran;Seo, Kyoung-Won;Song, Kun-Ho
    • Parasites, Hosts and Diseases
    • /
    • v.52 no.6
    • /
    • pp.691-694
    • /
    • 2014
  • The purpose of this study was to conduct a survey of Dirofilaria immitis infection among stray cats in Korea using nested PCR. We included 235 stray cats (121 females and 114 males) and evaluated each for the presence of feline heartworm infection. Blood samples were collected from 135 cats in Daejeon, 50 cats in Seoul, and 50 cats from Gyeonggi-do (Province). Of the 235 DNA samples, 14 (6.0%) were positive for D. immitis. The prevalence of infection in male cats (8/114, 7.0%) tended to be higher than that in female cats (6/121, 5.0%), but the difference was not statistically significant. In each location, 8, 2, and 4 cats were positive for infection, respectively, based on DNA testing. No significant differences in the prevalence were observed among the geographic regions, although the rate of infection was higher in Gyeonggi-do (8.0%) than Daejeon (5.9%) and Seoul (4.0%). We submitted 7 of the 14 D. immitis DNA-positive samples for sequencing analysis. All samples corresponded to partial D. immitis cytochrome c oxidase subunit I gene sequences with 99% homology to the D. immitis sequence deposited in GenBank (accession no. FN391553). To the best of our knowledge, this is the first survey using nested PCR to analyze the prevalence of D. immitis in stray cats in Korea.

Evidence-based Screening, Early Diagnosis and Treatment Strategy of Cervical Cancer for National Policy in Low-resource countries: Example of India

  • Saxena, Upma;Sauvaget, Catherine;Sankaranarayanan, Rengaswamy
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.4
    • /
    • pp.1699-1703
    • /
    • 2012
  • Cervical cancer remains the most frequent cancer in women from the developing world. More than 88% of deaths occur in low-income countries, and it is predicted to climb to 91.5% by 2030. Although Pap-based screening programmes have shown to be effective in reducing the disease burden in high-resource countries, implementation and sustention of cytology-based programmes is quite challenging in low-resource settings. The present paper reviews evidence-based alternatives of screening methods, triaging algorithm, treatment of cervical precancerous lesions, and age-group at screening appropriate for low-income countries. Evidence shows that visual inspection methods using diluted acid acetic or Lugol's iodine, and HPV-DNA testing are more sensitive tests than the Pap-smear screening test. Visual inspection allows an immediate result and, when appropriate, may be immediately followed by cryotherapy, the so called "screen-and-treat" approach, achieved in a single visit, by trained nurses and midwives. Examples of cervical cancer prevention programmes in India and selected low-income countries are given.

Statistical Tests for Time Course Microarray Experiments

  • Park, Tae-Seong;Lee, Seong-Gon;Choe, Ho-Sik;Lee, Seung-Yeon;Lee, Yong-Seong
    • Proceedings of the Korean Statistical Society Conference
    • /
    • 2002.05a
    • /
    • pp.85-90
    • /
    • 2002
  • Microarray technology allows the monitoring of expression levels for thousands of genes simultaneously. In time-course experiments in which gene expression is monitored over time we are interested in testing gene expression profiles for different experimental groups. We propose a statistical test based on the ANOVA model to identify genes that have different gene expression profiles among experimental groups in time-course experiments. Using this test, we can detect genes that have different gene expression profiles among experimental groups. The proposed model is illustrated using cDNA microarrays of 3,840 genes obtained in an experiment to search for changes in gene expression profiles during neuronal differentiation of cortical stem cells.

  • PDF